Liposomal particles, methods of making same and uses thereof
Liposomes termed as small unilamellar vesicles (SUVs), can be synthesized in the 20-50 nm size range, but encounter challenges such as instability and aggregation leading to inter-particle fusion. This limits their use as a therapeutic delivery agent. Increasing the surface negative charge of SUVs, via the attachment of anionic entities such as DNA/RNA, increases the colloidal stability of these vesicles. Additionally, the dense spherical arrangement and radial orientation of nucleic acids exhibits unique chemical and biological properties, unlike their linear counterparts. These liposomal particles, are non-toxic and though anionic, can efficiently enter cells without the aid of ancillary cationic transfection agents in a non-immunogenic fashion. These exceptional properties allow their use as delivery agents for gene regulation in different therapies and offer an alternative platform to metal core spherical nucleic acids.
1. A liposomal particle having a substantially spherical geometry, comprising:
a lipid bilayer comprising a plurality of lipid groups; and single-stranded oligonucleotides that are inhibitory RNA or antisense DNA, wherein all the oligonucleotides are attached to the surface of the liposomal particle, wherein each oligonucleotide is an oligonucleotide-lipid conjugate containing one lipophilic tethered group anchored into the external side of the lipid bilayer, wherein one or more of the oligonucleotides hybridize to a gene comprising a polynucleotide encoding a gene product and are complementary to all or a portion of the polynucleotide, wherein the hybridizing between the oligonucleotide and the polynucleotide occurs over a length of the polynucleotide with a degree of complementarity sufficient to inhibit expression of the gene product, and wherein the diameter of the liposomal particle is less than or equal to 50 nanometers (nm).
2. The liposomal particle of claim 1 , wherein the plurality of lipid groups comprises a lipid selected from the group consisting of the phosphatidylcholine, phosphatidylglycerol, and phosphatidylethanolamine family of lipids.
3. The liposomal particle of claim 2 , wherein the lipid is selected from the group consisting of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dimyristoyl-sn-phosphatidylcholine (DMPC), l-palmitoyl-2-oleoyl-sn-phosphatidylcholine (POPC), 1,2-distearoyl-sn-gl ycero-3-phospho-(l′-rac-glycerol) (DSPG), 1,2-dioleoyl-sn-glycero-3-phospho-(I′-rac-glycerol) (DOPG), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphoethanolamine (DOPE), and 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (DPPE).
4. The liposomal particle of claim 1 , wherein the lipophilic tethered group comprises a tocopherol or cholesterol.
5. The liposomal particle of claim 4 , wherein the tocopherol is chosen from the group consisting of a tocopherol derivative, alpha-tocopherol, beta-tocopherol, gam ma-tocopherol and delta-tocopherol.
6. The liposomal particle of claim 1 , wherein the inhibitory RNA is selected from the group consisting of a small inhibitory RNA (siRNA), a miRNA, an inhibitory RNA that forms a triplex with double stranded DNA, and a ribozyme.
7. The liposomal particle of claim 1 , wherein the liposomal particle comprises from about 10 to about 80 oligonucleotides.
8. The liposomal particle of claim 7 , wherein the particle comprises 70 oligonucleotides.
9. The liposomal particle of claim 7 , wherein the liposomal particle comprises 30 oligonucleotides.
10. The liposomal particle of claim 1 , wherein the oligonucleotides are modified oligonucleotides having phosphorothioate linkages.
11. The liposomal particle of claim 1 , wherein the diameter of the liposomal particle is less than or equal to 40 nanometers (nm).
12. The liposomal particle of claim 1 , wherein the diameter of the liposomal particle is less than or equal to 35 nanometers (nm).
13. The liposomal particle of claim 1 , wherein the diameter of the liposomal particle is less than or equal to 30 nanometers (nm).
14. The liposomal particle of claim 1 , wherein the lipophilic tethered group is at the 3′-end of each oligonucleotide.
15. A plurality of liposomal particles comprising a plurality of the liposomal particle of claim 1 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 50 nanometers (nm).
16. The plurality of liposomal particles of claim 15 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 40 nanometers (nm).
17. The plurality of liposomal particles of claim 15 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 35 nanometers (nm).
18. The plurality of liposomal particles of claim 15 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 30 nanometers (nm).
19. A liposomal particle having a substantially spherical geometry, consisting of:
a lipid bilayer consisting of a plurality of lipid groups; and single-stranded oligonucleotides that are inhibitory RNA or antisense DNA, wherein all the oligonucleotides are attached to the surface of the liposomal particle, wherein each oligonucleotide is an oligonucleotide-lipid conjugate containing one lipophilic tethered group anchored into the external side of the lipid bilayer, wherein one or more of the oligonucleotides hybridize to a gene comprising a polynucleotide encoding a gene product and are complementary to all or a portion of the polynucleotide, wherein the hybridizing between the polynucleotide and the oligonucleotide occurs over a length of the polynucleotide with a degree of complementarity sufficient to inhibit expression of the gene product, and wherein the diameter of the liposomal particle is less than or equal to 50 nanometers (nm).
20. A plurality of liposomal particles comprising a plurality of the liposomal particle of claim 19 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 50 nanometers (nm).
21. The plurality of liposomal particles of claim 20 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 40 nanometers (nm).
22. The plurality of liposomal particles of claim 20 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 35 nanometers (nm).
23. The plurality of liposomal particles of claim 20 , wherein the plurality of liposomal particles has a mean diameter of less than or equal to about 30 nanometers (nm).